Quantification of ZnO nanoparticle uptake, distribution, and dissolution within individual human macrophages.

The usefulness of zinc oxide (ZnO) nanoparticles has led to their wide distribution in consumer products, despite only a limited understanding of how this nanomaterial behaves within biological systems. From a nanotoxicological viewpoint the interaction(s) of ZnO nanoparticles with cells of the immune system is of specific interest, as these nanostructures are readily phagocytosed. In this study, rapid scanning X-ray fluorescence microscopy was used to assay the number ZnO nanoparticles associated with ∼1000 individual THP-1 monocyte-derived human macrophages. These data showed that nanoparticle-treated cells endured a 400% elevation in total Zn levels, 13-fold greater than the increase observed when incubated in the presence of an equitoxic concentration of ZnCl2. Even after excluding the contribution of internalized nanoparticles, Zn levels in nanoparticle treated cells were raised ∼200% above basal levels. As dissolution of ZnO nanoparticles is critical to their cytotoxic response, we utilized a strategy combining ion beam milling, X-ray fluorescence and scanning electron microscopy to directly probe the distribution and composition of ZnO nanoparticles throughout the cellular interior. This study demonstrated that correlative photon and ion beam imaging techniques can provide both high-resolution and statistically powerful information on the biology of metal oxide nanoparticles at the single-cell level. Our approach promises ready application to broader studies of phenomena at the interface of nanotechnology and biology.

Focused ion beam milling of microchannels in lithium niobate.

We present experimental and simulation results for focused ion beam (FIB) milling of microchannels in lithium niobate in this paper. We investigate two different cuts of lithium niobate, Y- and Z-cuts, and observe that the experimental material removal rate in the FIB for both Y-cut and Z-cut samples was 0.3 µm(3)/nC, roughly two times greater than the material removal rate previously reported in the literature but in good agreement with the value we obtain from stopping and range of ions in matter (SRIM) simulations. Further, we investigate the FIB milling rate and resultant cross-sectional profile of microchannels at various ion beam currents and find that the milling rate decreases as a function of ion dose and correspondingly, the cross-sectional profiles change from rectangular to V-shaped. This indicates that material redeposition plays an important role at high ion dose or equivalently, high aspect ratio. We find that the experimental material removal rate decreases as a function of aspect ratio of the milled structures, in good agreement with our simulation results at low aspect ratio and in good agreement with the material removal rates previously reported in the literature at high aspect ratios. Our results show that it is indeed easier than previously assumed to fabricate nanochannels with low aspect ratio directly on lithium niobate using the FIB milling technique.

Experimental characterization of a metal-oxide-semiconductor field-effect transistor-based Coulter counter.

We report the detailed characterization of an ultrasensitive microfluidic device used to detect the translocation of small particles through a sensing microchannel. The device connects a fluidic circuit to the gate of a metal-oxide-semiconductor field-effect transistor (MOSFET) and detects particles by monitoring the MOSFET drain current modulation instead of the modulation in the ionic current through the sensing channel. The minimum volume ratio of the particle to the sensing channel detected is 0.006%, which is about ten times smaller than the lowest detected volume ratio previously reported in the literature. This volume ratio is detected at a noise level of about 0.6% of the baseline MOSFET drain current, clearly showing the amplification effects from the fluidic circuits and the MOSFETs. We characterize the device sensitivity as a function of the MOSFET gate potential and show that its sensitivity is higher when the MOSFET is operating below its threshold gate voltage than when it is operating above the threshold voltage. In addition, we demonstrate that the device sensitivity linearly increases with the applied electrical bias across the fluidic circuit. Finally, we show that polystyrene beads and glass beads with similar sizes can be distinguished from each other based on their different translocation times, and the size distribution of microbeads can be obtained with accuracy comparable to that of direct scanning electron microscopy measurements.